Nanometer nucleic acid guide medicine, and preparation method and application thereof
A nucleic acid drug and drug technology, used in drug combinations, pharmaceutical formulations, anti-tumor drugs, etc., can solve the problems of inability to effectively reach the lesions, reduce the clinical application effect, and limit the effect of nucleic acid drugs.
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Embodiment 1
[0073] The synthesis of embodiment 1, C32
[0074] 1. Melt 400ul of 5-amino-1-pentanol in a 40°C water bath;
[0075] 2. Use a pipette to absorb 356ul of 5-amino-1-pentanol and 651ul of 1,4-butanediol diacrylate, 5-amino-1-pentanol and 1,4-butanediol diacrylate The molar ratio is 1.2:1.0, mix thoroughly and put into the centrifuge tube;
[0076] 3. Let it react at 94°C for 24 hours, and polymerize into C32 under suitable conditions;
[0077] 4. Dissolve the C32 completed in the above reaction with DMSO to prepare a 10ml solution (concentration is 100mg / ml);
[0078] 5. Sterile filter the solution with a 0.22um filter membrane, and distribute it in ten small centrifuge tubes of 1.5ml;
[0079] 6. Store at -20°C for later use.
Embodiment 2
[0080] Embodiment 2, MicroRNA
[0081] See Table 1 for valid nucleic acid chains. Including: Mir-34, miR-122, miR-125, miR-130a, miR-150, miR-199, miR-200 and let-7 family members.
[0082] Table 1
[0083]
Embodiment 3
[0084] Embodiment 3, the preparation of the nucleic acid medicine that comprises targeting peptide
[0085] Targeting polypeptides used (see Lee, et al, cancer research 2007:67(22), Jia et al Cancer Letters 247(2007) 234-242; Du et al, Mol Cancer Res.2010; 8(2):135-144 ) see Table 2.
[0086] Table 2
[0087] Peptide name
SEQ ID NO:
amino acid sequence
SP5-52
11
SVSVGMKPSPRP
12
AWYPLPP
13
AQYPLPP
A54
14
AGKGTPSLETTP
[0088] (a) C32-A54 targeting peptide modification
[0089]Add A54 (AGKGTPSLETTP) with a molar ratio of 1.0: 2.4 (C32: A54) in 5 milliliters of DMSO solution containing 1.9 grams of C32 to carry out esterification reaction. The optimum process condition for the synthesis reaction is: the molar ratio of C32 and A54 is 1.0 : 2.4, Catalyst SO 4 2- / SiO 2 The dosage is 0.19g, the reaction temperature is 60°C, and the reaction time is 4h. At this time, the esterification yield i...
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